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Deferoxamine Treatment Prevents Post-Stroke Vasoregression and Neurovascular Unit Remodeling Leading to Improved Functional Outcomes in Type 2 Male Diabetic Rats: Role of Endothelial Ferroptosis

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Abstract

It is a clinically well-established fact that patients with diabetes have very poor stroke outcomes. Yet, the underlying mechanisms remain largely unknown. Our previous studies showed that male diabetic animals show greater hemorrhagic transformation (HT), profound loss of cerebral vasculature in the recovery period, and poor sensorimotor and cognitive outcomes after ischemic stroke. This study aimed to determine the impact of iron chelation with deferoxamine (DFX) on (1) cerebral vascularization patterns and (2) functional outcomes after stroke in control and diabetic rats. After 8 weeks of type 2 diabetes induced by a combination of high-fat diet and low-dose streptozotocin, male control and diabetic animals were subjected to thromboembolic middle cerebral artery occlusion (MCAO) and randomized to vehicle, DFX, or tPA/DFX and followed for 14 days with behavioral tests. Vascular indices (vascular volume and surface area), neurovascular remodeling (AQP4 polarity), and microglia activation were measured. Brain microvascular endothelial cells (BMVEC) from control and diabetic animals were evaluated for the impact of DFX on ferroptotic cell death. DFX treatment prevented vasoregression and microglia activation while improving AQP4 polarity as well as blood-brain barrier permeability by day 14 in diabetic rats. These pathological changes were associated with improvement of functional outcomes. In control rats, DFX did not have an effect. Iron increased markers of ferroptosis and lipid reactive oxygen species (ROS) to a greater extent in BMVECs from diabetic animals, and this was prevented by DFX. These results strongly suggest that (1) HT impacts post-stroke vascularization patterns and recovery responses in diabetes, (2) treatment of bleeding with iron chelation has differential effects on outcomes in comorbid disease conditions, and (3) iron chelation and possibly inhibition of ferroptosis may provide a novel disease-modifying therapeutic strategy in the prevention of post-stroke cognitive impairment in diabetes.

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Acknowledgments

The authors would like to thank the summer medical students David Foreman and Thomas Menk for their assistance with the analysis of vascularization data with the Volocity software.

Funding

This study was supported by Veterans Affairs (VA) Merit Review (BX000347), VA Senior Research Career Scientist Award (IK6 BX004471), National Institute of Health (NIH) R01 NS083559 and R01 NS104573 (multi-PI, Susan C. Fagan as co-PI) to Adviye Ergul; Diabetic Complications Research Consortium DiaComp awards 17AU3831/18AU3903 (DK076169/115255) to Dr. Weiguo Li; Scientist Development Grant (16SDG30270013) and Mercer University seed grant to Dr. Mohammed Abdelsaid; and NIH T32 HL007260 to Victoria Wolf.

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  1. Yasir Abdul and Weiguo Li contributed equally to this work.

Authors and Affiliations

  1. Ralph H. Johnson VA Medical Center, Charleston, SC, USA

    Yasir Abdul, Weiguo Li, Sarah Jamil, Victoria Wolf & Adviye Ergul

  2. Department of Pathology and Laboratory Sciences, Medical University of South Carolina, 171 Ashley Ave. MSC 908, Charleston, SC, 29425, USA

    Yasir Abdul, Weiguo Li, Sarah Jamil, Victoria Wolf & Adviye Ergul

  3. Department of Medicine, Massachusetts General Hospital, Boston, MA, USA

    Rebecca Ward

  4. School of Medicine, Mercer University, Savannah, GA, USA

    Mohammed Abdelsaid

  5. Department of Pharmaceutical Sciences, College of Pharmacy, Larkin University, Miami, FL, USA

    Sherif Hafez

  6. Department of Neuroscience and Regenerative Medicine, Medical College of Georgia, Augusta, GA, USA

    Guangkuo Dong & Maribeth H. Johnson

  7. Program in Clinical and Experimental Therapeutics, University of Georgia College of Pharmacy, Augusta, GA, USA

    Susan C. Fagan

  8. Charlie Norwood Veterans Affairs Medical Center, Augusta, GA, USA

    Susan C. Fagan

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  1. Yasir Abdul
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All rats were housed in the animal care facility at Augusta University, which is approved by the American Association for Accreditation of Laboratory Animal Care. All experiments were conducted in accordance with the National Institutes of Health (NIH) guidelines for the care and use of animals in research. Furthermore, all protocols were approved by the institutional animal care and use committee.

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Abdul, Y., Li, W., Ward, R. et al. Deferoxamine Treatment Prevents Post-Stroke Vasoregression and Neurovascular Unit Remodeling Leading to Improved Functional Outcomes in Type 2 Male Diabetic Rats: Role of Endothelial Ferroptosis. Transl. Stroke Res. 12, 615–630 (2021). https://doi.org/10.1007/s12975-020-00844-7

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